Development and validation of a quantitative PCR assay for detection of Sulawesi tortoise adenovirus.

In 2007, a mortality event involving over 100 Sulawesi tortoises (Indotestudo forsteni), two Impressed tortoises (Manouria impress) and a critically endangered Burmese star tortoise (Geochelone platynota) was attributed to Sulawesi tortoise adenovirus (STADV; genus Siadenovirus). We developed a TaqMan quantitative PCR assay targeting the DNA polymerase gene of STADV for use in clinical diagnosis and epidemiologic surveillance. This assay failed to amplify five closely-related chelonian adenoviruses, indicating high analytical specificity. The assay performed with high efficiency (slope = -3.337; R2 = 0.999) and high inter- and intra-assay repeatability (coefficient of variation <1.36 % at all standard curve dilutions). Dynamic range included 1.00 × 107 to 1.00 × 101 target copies per reaction and limit of detection was 101 target copies per reaction, though 100 target copies per reaction were intermittently detected. This qPCR assay provides a valuable diagnostic tool for characterization of STADV epidemiology, including potential identification of the North American reservoir host.

Parasites of reptiles in Iran (1922-2023): A literature review.

Reptiles are among the most diverse groups of animals, inhabiting nearly all continents and environments. Understanding their parasite biodiversity has garnered significant interest, particularly from a One Health perspective. Although the highly diverse reptile fauna of Iran, comprising 272 species i.e. 89 snakes (Serpentes), 171 lizards (Sauria), 8 turtles, 2 tortoises (Testudines), 1 crocodile (Crocodilia), and 1 worm-lizard (Amphisbaenia), there is a shortage of information about parasites. The present review is a compilation of 62 studies published from 1922 to August 2024. We present information on 56 species of reptiles from five groups (amphisbaenians, crocodiles, testudines, snakes, and lizards) and 98 parasitic taxa belonging to different protozoa and metazoa i.e. nematodes, cestodes, trematodes, acanthocephala, leeches, ticks, mites, and myiasis -producing flies. Although 63 taxa were diagnosed at the species level, 35 parasite taxa were only reported at the genus or family levels. Reviewing the literature, we found a paucity of information about endemic reptiles several of which are vulnerable species. Considering that some of the detected parasites e.g. Cryptosporidium and amoebae have serious clinical and/or public health threats molecular diagnostic techniques are needed for precise identification and understanding of the epidemiology and the potential zoonotic implications associated with parasites of reptiles. There is also a need to understand the exact distribution and host-parasite associations in different reptilian species present in Iran including the role of the reptiles as intermediate and reservoir hosts.

Molecular detection and phylogenetic characterization of Rickettsia in ticks collected from leopard tortoise (Geochelone pardalis) in rural Zambia.

In sub-Saharan Africa, limited studies have investigated zoonotic pathogens that may be harboured by ticks infesting reptiles such as tortoises. Here, we report the presence of pathogenic Rickettsia in ticks (Amblyomma marmoreum) collected from the leopard tortoise (Geochelone pardalis) in rural Zambia. Using polymerase chain reaction, 56% (49/87) of ticks were positive for the Rickettsia outer membrane protein (ompB) gene. Multi-locus sequence and phylogenetic analysis based on the ompB, ompA, and citrate synthase (gltA) genes showed that the ticks carried R. africae, and other Rickettsia spp. closely related to R . raoultii, R . massiliae, R . tamurae and R . monacensis. Given the proximity between humans, livestock, and wildlife in these habitats, there exists a considerable risk of transmission of zoonotic Rickettsia to human populations in this rural setting. These results call for heightened awareness and further research into the dynamics of tick-borne diseases in regions where humans and animals coexist, particularly in the context of tortoise-associated ticks as vectors. Understanding and addressing these potential disease vectors is crucial for effective public health measures and the prevention of Rickettsia zoonoses.

Trace Elements in Hermann's Tortoises (Testudo hermanni) According to Sex, Season, and Sampling Region in Central Europe.

Some trace elements are essential for the health of vertebrates, but little is known about their function, the amounts required, and the factors influencing their metabolism in tortoises. The aim of this study was therefore to measure trace elements (chromium (Cr), cobalt (Co), copper (Cu), iron (Fe), magnesium (Mg), manganese (Mn), molybdenum (Mo), selenium (Se), zinc (Zn)) in heparinized blood plasma of Hermann's tortoises (Testudo hermanni) (n = 520) from March to September 2022 using inductively coupled plasma mass spectrometry (ICP-MS) and to establish specific reference intervals. Additionally, the influence of sex, season, and region of sample collection on the measured values were evaluated. Significant (p ≤ 0.05) sex-specific differences were found for Cu, Mg, and Mn; seasonal differences were found for Cr, Cu, Mn, Mo, and Se; and the region in which the tortoises were kept significantly impacted Cr, Cu, Fe, Mg, Mo, and Se levels. The results show that all of these factors should be consider when establishing and interpreting blood trace element levels in tortoises.

An efficient model for extracting respiratory and blood oxygen saturation data from photoplethysmogram signals by removing motion artifacts using heuristic-aided ensemble learning model.

Patients with surgical, pulmonary, and cardiac problems, continual monitoring of Oxygen Saturation of a Person (SpO2) and Respiratory Rate (RR) is essential. Similarly, the persons with cardiopulmonary health issues, RR estimation is crucial. The performance of the ventilator assistance and lung medicines are evaluated using SpO2 and RR. For the persons, those who are living alone with respiratory illnesses need a compulsory estimation of RR. In case of serious illness, the RR might face abrupt changes. The immobility of the disturbance and RR makes the RR evaluation from the PhotoPlethysmoGraphic (PPG) signals is a difficult challenge. So, an efficient RR and SpO2 estimation framework from the PPG signal using the deep learning method is developed in this paper. At first, the PPG signal is collected from standard data sources. The collected PPG signals undergo signal pre-processing. The pre-processing procedures include Motion Artifacts (MA) removal and filtering techniques. The pre-processed signals are split into distinct windows. From the split windows of the signals, the spectral features, RR, and Respiratory Peak Variance (RPV) features are extracted. The retrieved features are selected optimally with the help of Advanced Golden Tortoise Beetle Optimizer (AGTBO). The weights are chosen optimally with the same AGTBO. The optimally selected features are fused with the optimal features to get the weighted optimal features. These weighted optimal features are fed into the Ensemble Learning-based RR and SpO2 Estimation Network (ELRR-SpO2EN). The ensemble learning model is developed by combining Multilayer Perceptron (MLP), AdaBoost, and Attention-based Long Short Term Memory (A-LSTM). The performance of the developed RR and SpO2 estimation model is compared with other existing techniques. The experimental analysis results revealed that the proposed AGTBO-ELRR-SpO2EN model attained 96 % accuracy for the second dataset, which is higher than the conventional models such as MLP (90 %), Adaboost (92 %), A-LSTM (92 %), and MLP-ADA-ALSTM (94 %). Thus, it has been confirmed that the designed RR and SpO2 estimation framework from PPG signals is more efficient than the other conventional models.

A novel immunofluorescence study of Lingual Salivary Glands in the Egyptian Tortoise (Testudo kleinmanni) and its ecological significance.

The Egyptian tortoise (Testudo kleinmanni) is remarkably adapted to its harsh desert environment, a characteristic that is crucial for its survival under extreme conditions. This study was aimed at providing a deeper understanding of the lingual salivary gland structures in the Egyptian tortoise and examining how these structures help the tortoise manage hydration and nutrition in arid conditions. Utilizing a combination of light microscopy and immunofluorescence, this research introduced pioneering methods involving seven different antibodies, marking a first in the study of reptilian salivary glands. Our investigations categorized the tortoise's salivary glands into papillary and non-papillary types. The papillary glands were further classified into superficial, deep, interpapillary, and intraepithelial salivary glands, while non-papillary glands included superficial and deep lingual types. Structurally, these glands are organized into lobules, delineated by interlobular septa, and are equipped with a duct system comprising interlobular, intercalated, and main excretory ducts with gland openings on the tongue's surface and the papillae surfaces. Notably, the superficial glands displayed both tubuloalveolar and acinar configurations, whereas the deep lingual glands were exclusively acinar. Immunofluorescence results indicated that α-smooth muscle actin (α-SMA) was prevalent in myoepithelial cells, myofibroblasts, and blood vessels, suggesting their integral role in glandular function and support. E-cadherin was predominantly found in epithelial cells, enhancing cell adhesion and integrity, which are critical for efficient saliva secretion. Importantly, Mucin 1 (MUC1) and Mucin 5B (MUC5B) staining revealed that most glands were mucous in nature, with MUC5B specifically marking mucin within secretory cells, confirming their primary function in mucous secretion. PDGFRα and CD34 highlighted the presence of telocytes and stromal cells within the glandular and interlobular septa, indicating a role in structural organization and possibly in regenerative processes. Cytokeratin 14 expression was noted in the basal cells of the glands, underscoring its role in upholding the structural foundation of the epithelial barrier. In conclusion, this detailed morphological and immunological characterization of the Egyptian tortoise's salivary glands provides new insights into their complex structure and essential functions. These findings not only enhance our understanding of reptilian physiology but also underline the critical nature of salivary glands in supporting life in arid environments. This study's innovative use of a broad range of immunofluorescence markers opens new avenues for further research into the adaptive mechanisms of reptiles.

An intelligent wireless channel corrupted image-denoising framework using symmetric convolution-based heuristic assisted residual attention network.

Image denoising is one of the significant approaches for extracting valuable information in the required images without any errors. During the process of image transmission in the wireless medium, a wide variety of noise is presented to affect the image quality. For efficient analysis, an effective denoising approach is needed to enhance the quality of the images. The main scope of this research paper is to correct errors and remove the effects of channel degradation. A corrupted image denoising approach is developed in wireless channels to eliminate the bugs. The required images are gathered from wireless channels at the receiver end. Initially, the collected images are decomposed into several regions using Adaptive Lifting Wavelet Transform (ALWT) and then the "Symmetric Convolution-based Residual Attention Network (SC-RAN)" is employed, where the residual images are obtained by separating the clean image from the noisy images. The parameters present are optimized using Hybrid Energy Golden Tortoise Beetle Optimizer (HEGTBO) to maximize efficiency. The image denoising is performed over the obtained residual images and noisy images to get the final denoised images. The numerical findings of the developed model attain 31.69% regarding PSNR metrics. Thus, the analysis of the developed model shows significant improvement.

Conservation units alone are insufficient to protect Brazilian Amazonian chelonians.

The creation of protected areas (PAs) is not always based on science; consequently, some aquatic species may not receive the same level of protection as terrestrial ones. The objective of this study was to identify priority areas for the conservation of chelonians in the Brazilian Amazon basin and assess the contribution of PAs, distinguishing between Full Protection Areas, Sustainable Use Areas, and Indigenous Lands for group protection. The entire species modeling procedure was carried out using Species Distribution Models. Location records were obtained from platforms such as SpeciesLink, GBIF, the Hydroatlas database, and WorldClim for bioclimatic variables adjusted with algorithms like Maximum Entropy, Random Forest, Support Vector Machine, and Gaussian-Bayesian. Indigenous lands cover more than 50% of the distribution areas of chelonian species in the Brazilian Amazon. Protected areas with higher conservation importance (Full Protection Areas and Sustainable Use Areas) hold less than 15% of the combined species distribution. Researchers face significant challenges when making decisions with models, especially in conservation efforts involving diverse taxa that differ significantly from one another within a group of individuals.

Macroscopic, microscopic, and immunofluorescent characterization of the Greek tortoise (Testudo graeca graeca) oropharyngeal floor with concern to its feed adaptation as a herbivorous land reptile.

The current investigation focuses on gross anatomy, light, and scanning electron microscopy (SEM) of the Testudo graeca oropharyngeal floor, with particular reference to the immunofluorescence technique to examine its tongue. The T. graeca oropharyngeal floor showed many anatomical structures: the lower rhamphotheca, paralingual ridge, lower alveolar ridge, tongue, laryngeal mound, and glottis. The lower rhamphotheca appeared as a V-shaped jaw line with a highly serrated edge and a median tomium (beak). SEM observations of the lingual apex and the lingual body showed rectangular and conical filiform papillae with porous surfaces and taste pores. Meanwhile, the lingual root had two wings that carried papillae with different shapes: dagger-shaped, conical, bifurcated, and leaf-like papillae, and these papillae lacked taste pores. The laryngeal mound had openings for the laryngeal mucus gland and its secretions. Light microscopy findings showed mucous glands in the propria submucosa and near the mucosal surface of the lingual apex. The lingual root had lingual papillae and two hyaline cartilaginous skeletons between skeletal muscles, and the lingual papillae were elongated filiform, rectangular filiform papillae, and fungiform papillae. The lamina propria constituted the core of the lingual papillae and the mucous gland, they had a positive reaction with the periodic acid schiff (PAS) reagent. The apical surface of the fungiform papillae had taste pores. Under immunofluorescence, the vimentin was detected in taste bud cells, and synaptophysin reacted to the taste buds and nerve bundles. The current study of the Greek tortoise oropharyngeal floor investigated its herbivorous eating habits using its serrated lower rhamphotheca, a large tongue with differently shaped papillae, and numerous mucous glands. RESEARCH HIGHLIGHTS: The Greek tortoise (T. graeca graeca) oropharyngeal floor showed many anatomical structures: lower rhamphotheca, paralingual ridge, lower alveolar ridge, tongue, laryngeal mound, and glottis. SEM and light microscopy observations of the tongue revealed varied types and shapes of lingual papillae with a porous surface on the tongue apex (rectangular or conical filiform papillae), on the tongue body (filiform and fungiform papillae), and on the tongue root (dagger-shaped, conical, bifurcated, and leaf-like papillae). Light microscopy findings: the lamina propria constituted the core of the lingual papillae and had numerous mucous glands that had a slightly magenta-red color with PAS reagent. The apical surface of the fungiform papillae had taste pores. Vimentin and synaptophysin gave a reaction to the taste buds.

Extracellular symbiont colonizes insect during embryo development.

Insects typically acquire their beneficial microbes early in development. Endosymbionts housed intracellularly are commonly integrated during oogenesis or embryogenesis, whereas extracellular microbes are only known to be acquired after hatching by immature instars such as larvae or nymphs. Here, however, we report on an extracellular symbiont that colonizes its host during embryo development. Tortoise beetles (Chrysomelidae: Cassidinae) host their digestive bacterial symbiont Stammera extracellularly within foregut symbiotic organs and in ovary-associated glands to ensure its vertical transmission. We outline the initial stages of symbiont colonization and observe that although the foregut symbiotic organs develop 3 days prior to larval emergence, they remain empty until the final 24 h of embryo development. Infection by Stammera occurs during that timeframe and prior to hatching. By experimentally manipulating symbiont availability to embryos in the egg, we describe a 12-h developmental window governing colonization by Stammera. Symbiotic organs form normally in aposymbiotic larvae, demonstrating that these Stammera-bearing structures develop autonomously. In adults, the foregut symbiotic organs are already colonized following metamorphosis and host a stable Stammera population to facilitate folivory. The ovary-associated glands, however, initially lack Stammera. Symbiont abundance subsequently increases within these transmission organs, thereby ensuring sufficient titers at the onset of oviposition ~29 days following metamorphosis. Collectively, our findings reveal that Stammera colonization precedes larval emergence, where its proliferation is eventually decoupled in adult beetles to match the nutritional and reproductive requirements of its host.

Exploring Aeromonas dhakensis in Aldabra giant tortoises: a debut report and genetic characterization.

Aeromonas dhakensis (A. dhakensis) is becoming an emerging pathogen worldwide, with an increasingly significant role in animals and human health. It is a ubiquitous bacteria found in terrestrial and aquatic milieus. However, there have been few reports of reptile infections. In this study, a bacterial strain isolated from a dead Aldabra giant tortoise was identified as A. dhakensis HN-1 through clinical observation, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS), and gene sequencing analysis. Subsequently, to evaluate its pathogenicity, the detection of virulence genes and mice infection experiments were performed. A. dhakensis HN-1 was found to contain seven virulence genes, including alt, ela, lip, act, aerA, fla, and hlyA. Mice infected with A. dhakensis HN-1 exhibited hemorrhage of varying degrees in multiple organs. The half-maximal lethal dose (LD50) value of A. dhakensis HN-1 for mice was estimated to be 2.05 × 107 colony forming units (CFU)/mL. The antimicrobial susceptibility test revealed that A. dhakensis HN-1 was resistant to amoxicillin, penicillin, ampicillin and erythromycin. This is the first report of A. dhakensis in Aldabra giant tortoises, expanding the currently known host spectrum. Our findings emphasize the need for One Health surveillance and extensive research to reduce the spread of A. dhakensis across the environment, humans, and animals.

Are the ladybugs Cryptolaemus montrouzieri and Exochomus quadripustulatus (Coleoptera: Coccinellidae) candidate predators of Toumeyella parvicornis (Hemiptera: Coccidae)?

BACKGROUND: Toumeyella parvicornis is an alien pest of recent introduction in Italy that infests stone pines (Pinus pinea L.), its main host plant in Europe. Infestations are currently controlled through endotherapic treatments, but the high costs and the long-term inefficacy highlight the need for alternative control strategies applicable in natural systems as well. An analysis of the effectiveness of autochthonous and naturalized predators is the first step to achieve this goal. This work focused on two candidate predators, Exochomus quadripustulatus and Cryptolaemus montrouzieri (Coleoptera: Coccinellidae), that may potentially control T. parvicornis. The analysis was carried out under laboratory-controlled and semi-field conditions. RESULTS: Laboratory tests were carried out to assess: (i) the predation rate of the ladybugs over different densities of pest preimaginal stages, and (ii) the effect of the presence of heterospecific and conspecific predators on the attractiveness of prey colonies. Semi-field experimentations were carried out by wrapping infested stone pine twigs with net sleeves and applying one of three treatments: (i) Exochomus quadripustulatus, (ii) Cryptolaemus montrouzieri, and (iii) control containing only the pest population. Both the ladybugs were attracted by T. parvicornis, offered as prey. Exochomus quadripustulatus was more attracted to conspecific and heterospecific ladybugs than Cryptolaemus montrouzieri and responded to prey more quickly. CONCLUSIONS: The results obtained could be useful for planning further experimentations to explore the potential use of these predators in biological control programs that may be applied in infested areas. Besides the use in an urban context, where pesticide use is strongly limited, the release of natural enemies may help safeguard stone pine forests. © 2024 Society of Chemical Industry.

Effects of inhibition of Nav1.3, Nav1.7, and Nav1.8 channels on pain-related behavior in Speke's hinge-back tortoise (Kinixys spekii).

Comparative studies using reptiles as experimental animals in pain research could expand our knowledge on the evolution and adaptation of pain mechanisms. Currently, there are no data reported on the involvement of voltage-gated sodium ion channels on nociception in reptiles. The aim of this study was to investigate the involvement of Nav1.3, Nav1.7, and Nav1.8 ion channels in nociception in Speke's hinge-back tortoise. ICA 121341 (selective blocker for Nav1.1/Nav1.3), NAV 26 (selective blocker for Nav1.7), and A803467 (selective blocker for Nav1.8) were used to investigate the involvement of Nav1.3, Nav1.7, and Nav1.8, respectively. The chemicals were administered intracoelomically thirty minutes before the start of nociceptive tests. ICA 121341 did not cause a significant decrease in the time spent in pain-related behavior in all the nociceptive tests. NAV 26 and A8034667 caused a statistically significant decrease in the mean time spent in pain-related behavior in the formalin and capsaicin tests. Only A803467 caused a statistically significant increase in the mean latency to pain-related behavior in the hot plate test. NAV 26 and A803467 had no observable side effects. In conclusion, Nav1.7 and Nav1.8 are involved in the processing of chemically induced inflammatory pain in Speke's hinge back tortoise. In addition, Nav1.8 are also significantly involved in the development of thermal-induced pain-related behavior in this species of reptile. However, our results do not support the involvement of Nav1.3 on the development of chemical or thermal induced pain-related behavior in the Speke's hinge back tortoise.

The complete mitochondrial genome of the red-footed tortoise (Chelonoidis carbonarius Spix 1824).

In this study, we report the complete mitochondrial genome of the red-footed tortoise (Chelonoidis carbonarius). The compete mitochondrial genome of C. carbonarius is 16,639 bp in length, including 13 protein-coding genes, 22 tRNA genes, and two rRNA genes. The mitogenome was deposited in NCBI GenBank under the accession number OQ789392. Furthermore, we also constructed a phylogenetic tree of Chelonoidis using eight species. These results will aid the conservation of Chelonoidis from the perspective of genetic evolution.

Evaluation of changes in serum biochemical parameters in zarudny's spur-thighed tortoises (Testudo graeca zarudnyi) influenced by gender and season.

BACKGROUND: The Zarudny's spur-thighed tortoise or Iranian tortoise (Testudo graeca zarudnyi) has just been reported from Iran so far. Therefore, the purpose of this study was to determine the effects of season and gender on serum biochemical parameters of this valuable species in Iran. RESULTS: This study was performed on 20 clinically healthy adult Zarudny's spur-thighed tortoises. Blood samples were collected from the jugular vein and then serum biochemical parameters and body weight were measured in autumn and winter. The following biochemical parameters were measured: total cholesterol, triglyceride, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, very low-density lipoprotein, total protein, creatinine, urea, glucose, calcium, inorganic phosphorus, total bilirubin, uric acid, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and lactate dehydrogenase. Urea concentration increased significantly at emergence from hibernation. Also, in winter, total protein, phosphorous, creatinine, total bilirubin concentrations and alkaline phosphatase activity decreased significantly compared to autumn, but aspartate aminotransferase and lactate dehydrogenase activities were significantly higher than in autumn. There was no significant difference for the parameters mentioned above by gender, except for phosphorous. Phosphorous concentration in females was significantly higher than that of males in autumn. Cholesterol and low-density lipoprotein cholesterol levels in females were significantly higher than males in autumn and winter. CONCLUSIONS: The present study is the first study that monitors the serum biochemical parameters of adult Zarudny's spur-thighed tortoises based on season and gender. Seasonal reference intervals should be used for biochemical parameters in this valuable species. Also, sex-specific reference intervals for phosphate and cholesterol are necessary.

Description of larva, pupa, and genitalia of Hybosa acutangula Spaeth, 1913 (Coleoptera: Chrysomelidae: Cassidinae) from the Brazilian Cerrado

ABSTRACT Great advances were made in recent years regarding the description of immatures of Cassidinae and their taxonomy as a whole, but many taxa remain undescribed. This study focuses on updating morphological data for Hybosa acutangula Spaeth, 1913 (Chrysomelidae: Cassidinae: Ischyrosonychini), a tortoise beetle native of South America, which was collected in Fridericia florida (DC.) L. G. Lohmann in the Brazilian Cerrado. We provide descriptions and illustrations of the morphology of the larva, pupa, and genitalia of adults, all of which are novel for this genus. The main discerning features for this species among other Ischyrosonychini are the lack of dark patterns in the dorsum of either the larvae, being limited to the dark scoli and the cranium, or the pupae, and a much reduced anal fork. We also present the first record of parasitism by Chalcididae wasps, Brachymeria sp. Westwood, 1832 and Conura sp. Spinola, 1837. Morphological comparisons remain limited demanding further studies with other species of Ischyrosonychini, as to better understand the placement of this species within the taxonomy of tortoise beetles.

Tick infestation in spur-thighed tortoise population: a pilot study for unraveling epidemiological patterns and demographic consequences.

Ectoparasites, such as ticks, modulate host population dynamics by impacting demographic traits. They transmit infectious agents among their hosts, posing a critical threat to animal and public health. This study aimed to characterize and analyze the Hyalomma aegyptium infestation on one of its main hosts, the spur-thighed tortoise, its effects on demographic traits, and to determine the diversity of infectious agents present in both ticks and tortoises in the Maamora forest (northwestern Morocco). Our results show that 100% of the tortoises were parasitized by adult ticks in spring, an infestation intensity of 4 ticks/tortoise (5.1 and 3.6 ticks/tortoise in males and females, respectively; 4.2 and 3.3 ticks/tortoise in gravid and non-gravid females, respectively) and an abundance ranging from 1 to 12. Although without significant differences, male tortoises had higher tick abundances than females. The interaction of tortoise sex and body condition was significantly related to tick abundance, male body condition decreased with higher tick abundance in contrast to females. Nevertheless, the interaction of body condition and reproductive stage of females was not significantly related to tick abundance. Gravid females were significantly associated with tick abundance, showing a slightly higher infestation than non-gravid females. Molecular analysis of pooled tick samples revealed the presence of Ehrlichia ewingii, Candidatus Midichloria mitochondrii, and Rickettsia africae, with a minimum infection rate of 0.61 to 1.84%. However, blood sample analysis of the tortoises was infectious agent-free, pinpointing a lack of significant health problems. Given the possible effect on the transmission of zoonotic diseases by spur-thighed tortoises associated with their frequent collection as pets, it should be surveyed to control possible human health problems. In conservation terms, as a long-lived species, the role of tick infestation in demographic traits might be included in the management and conservation programs of spur-thighed tortoises.

New Recording of Toxoplasma gondii in Wild Tortoise Testudo graeca Using Nested PCR Method.

Toxoplasmosis is one of the most widespread zoonotic diseases, especially in warm and humid areas, and affects all mammals, including humans and many herbivores and carnivores. The present study investigated the Toxoplasma gondii (T. gondii) parasite in tortoises for the first time in Iraq using PCR technology. A total of 28 tortoises/Testudo graeca (T. graeca) were collected between October 2018 and March 2019 from the study stations and then sent to the Animal House, which belongs to the Department of Biology, Faculty of Education, University of AL-Qadisiyah, Iraq, to perform the dissection. The body cavity was opened, and all organs were removed. The tortoises' liver, heart, and brain were removed and kept at -20ºC until use. Afterward, the samples were subjected to DNA extraction. The Nested-PCR technique was implemented using two pairs of primers, and then the PCR products were analyzed using 1.5% agarose gel electrophoresis. The amplification of the gene during the first cycle indicated that 10 samples gave positive results with a total percentage of (11.9%), including five liver samples, three heart samples, and two brain samples (17.85%, 10.71%, and 7.14%, respectively). On the other hand, during the second cycle of the reaction, the amplification of the gene was obtained in seven samples (8.33%). The highest percentage of the presence of the gene was recorded in the tortoises' liver (14.28%) and the lowest in their brain (3.57%). This study is among the first to investigate the molecular detection of T. gondii in wild tortoises (T. graeca) in Iraq. The findings imply that tortoises have a role in transmitting T. gondii and are believed to acquire infection by feeding on small invertebrate animals or plants contaminated with the oocysts of the parasite.

Case report: Intracoelomic neoplastic mass of undetermined origin in an Asia minor spur-thighed tortoise (Testudo graeca ibera).

This article describes the diagnostic, treatment and attempted characterization of a neoplasia of undetermined origin in a Asia minor spur-thighed tortoise. A 21-year-old male Asia minor spur-thighed tortoise (Testudo graeca ibera) was admitted for a 4-month history of diarrhea, and a 2-month history of anorexia and lethargy. Physical examination revealed a firm midcoelomic mass in the right prefemoral fossa. Blood biochemistry indicated hypocalcemia and mild elevation of aspartate aminotransferase. Supportive care was administered in the form of heating, baths, and calcium injections. Ultrasound examination of the coelomic cavity revealed a 6-cm diameter, highly vascularized mass with liver-like echogenicity. Neoplasia was suspected, and endoscopy was performed, revealing a brown circumscribed mass with smooth edges. Surgical removal of the mass was evaluated by CT scan and achieved via a plastrotomy; however, the patient died 1 day post-surgery. The mass was located on the dorsal right side of the coelomic cavity in the anatomic location of the right testicle. Histopathology revealed neoplastic cells organized in packets supported by fibrous septa. Neoplastic cells showed moderate and inconsistent positive immunohistochemical labeling for S100 and NSE, and negative immunohistochemical labeling for pan-cytokeratin, vimentin, CD3, CD79a, chromogranin A, and synaptophysin. The prominent histological and anatomical characteristics of the mass indicated a possible testicular or neuroendocrine (e.g., adrenal gland) origin. Due to inconclusive immunohistochemical profiles and poorly differentiated neoplastic cells, only a final diagnosis of intracoelomic malignant tumor of undetermined origin could be established. This case underscores the difficulties encountered in achieving definitive diagnoses of neoplastic diseases in reptile medicine.

Fully 3D-printed tortoise-like soft mobile robot with muti-scenario adaptability.

Soft robotic systems are well suited to unstructured, dynamic tasks and environments, owing to their ability to adapt and conform without damaging themselves or their surroundings. These abilities are crucial in areas such as human-robot interaction, simplification of control system and weight reduction. At present, the existing soft mobile robots still have the disadvantages of single motion mode and application scenario, difficult manufacturing and low energy conversion efficiency. Based on the current shortcomings of soft robots, this paper designs and proposes a fully 3D-printed tortoise-like soft mobile robot with muti-scenarios adaptability. The robot uses a Bionic Tortoise Leg Actuator structure that enables simultaneous bending of the actuator in both directions, simplifying robot control and increasing the maximum bending angle achievable. In addition, a reconfiguration design solution has been proposed to enable the robot to implement two bionic modes for land and sea turtles, adapting to move on hard and soft surfaces and in water, enabling it to move in amphibious and complex environments. The performance of the pneumatic soft actuator is also improved by an improved Digital Light Processing method that enhances the maximum strain of the 3D printed soft material. The prototype was tested to give maximum movement speeds for different gaits and environments, demonstrating that the fully 3D printed tortoise-like soft-mobile robot designed in this paper is highly adaptable to multiple scenarios. The robot studied in this paper has a wide range of applications, with potential applications including navigation in a variety of domain environments, inspection of large underground oil and gas pipelines, and navigation in high temperature, high humidity and strong magnetic field environments or in military alert conditions.